摘要
目的:探讨p-p38 MAPK在腺苷A2A受体敲除(A2AR-/-)新生小鼠缺氧缺血脑区的表达及意义。方法:采用改良Rice法建立新生小鼠缺氧缺血性脑损伤(HIBD)模型。A2AR-/-(KO)及同期野生型(A2AR+/+,WT)C57/BL6新生小鼠(7日龄)分别按照完全随机分组方法分成假手术组及模型组,模型组按HIBD后取标本时间不同又分为造模后1 d组、3 d组和7 d组,共8个实验动物组,每组取小鼠8只,共64只。采用TUNEL结合尼氏染色检测神经细胞凋亡,采用免疫组织化学法检测活化caspase-3及p-p38 MAPK表达。同时,KO及同期WT小鼠分别取假手术组(SKO和SWT,n=10)及模型组(MKO和MWT,n=30)于HIBD后1 d同时进行新生鼠短期神经行为学评定。结果:(1)缺氧缺血后皮层及海马CA1区神经细胞凋亡、活化caspase-3及p-p38 MAPK表达均增加,造模后1 d为高峰;(2)A2AR敲除导致神经细胞凋亡及活化caspase-3表达增加,与同一时点的WT小鼠相比均有显著差异(P<0.01);p-p38 MAPK表达增加,其中,1 d及3 d后2种基因型小鼠间均有显著差异(P<0.01);(3)Pea-son相关分析显示,活化caspase-3表达与p-p38 MAPK表达呈显著正相关(皮层:r=0.957,P<0.01;海马CA1区:r=0.939,P<0.01)。结论:p-p38 MAPK可能参与了A2AR敲除增加新生小鼠脑缺氧缺血后神经细胞凋亡、加重脑损害的过程。
AIM： To study the expression of p-p38 MAPK in partial cerebral tissues after hypoxic-ischemic brain damage （HIBD） in the neonatal adenosine AEA receptor knockout （A2AR-/- ） mice. METHODS： Base on the modi- fied Rice method, the model of HIBD was established. The total 64 C57/BL6 neonatal mice （7 days old） of A24R^-/- （KO） and corresponding wild type （A24R^-/- , WT） were randomized into sham-operated group and model group. The mice in model group were divided into 3 subgroups ： I dO after HIBD, 3 d after HIBD and 7 d after HIBD （ n = 8 for each group）. The cortex and hippocampal CA1 region were used as the study areas. The neuronal apoptosis was detected using TUNEL assay combined with Nissl staining. The expression of p-p38 MAPK and activated caspase-3 was determined by the method of immunohistochemistry. The KO mice and WT mice were also taken from sham-operated group （ SKO and SWT, n = 10） and model group （MKO and MWT, n =30） 1 d after HIBD to assess the early neurological behavior. RESULTS： The apoptotic neurons, activated caspase-3 and p-p38 MAPK increased after HIBD and peaked at 1 d after HIBD in the cortex and the hippocampal CA1 region. The apoptotic neurons and the expression of activated caspase-3 in KO mice were significantly higher than those in WT mice at the same time point after HIBD. The expression level of p-p38 MAPK in KO mice were significantly higher than that in WT mice at 1 d and 3 d after HIBD. The expression of activated caspase-3 was positively correlated with the expression of p-p38 MAPK in neonatal mice after HIBD （ in the cortex： r = 0.957, P 〈 0. 01 ;in the hippoeampal CA1 region： r = 0.939, P 〈 0. 01 ）. CONCLUSION： p-p38 MAPK might be involved in the aggrava- ted neuron apoptosis and brain damage induced by AzAR knockout after neonatal HIBO.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第1期36-42,共7页
Chinese Journal of Pathophysiology
基金
浙江省科技厅钱江人才项目(No.2009R10024)
温州市科技局对外合作项目(No.H20110018)
